Human Embryonic and Hepatic Stem Cell Differentiation Visualized in Two and Three Dimensions Based on Serial Sections

Publikation: Bidrag til bog/antologi/rapportBidrag til bog/antologiForskningfagfællebedømt

Peter S. Vestentoft, Christian B Brøchner, Niels Lynnerup, Claus Yding Andersen, Kjeld Møllgård

Pluripotent human embryonic stem cells (hESCs) are characterized by two defining properties, self-renewal and differentiation. Self-renewing hESCs express transcription factors OCT4, SOX2, and NANOG, and surface markers SSEA-4 and TRA-1-60 and TRA-1-81 and their ability to differentiate into derivatives of the three germ layers show the differentiating potential. Studies suggest a certain microheterogeneity of the hESC colonies, in which not all cells in one colony of apparently undifferentiated cells express all the expected markers. We describe a technique to paraffin embed an entire hESC colony, and prepare 3-5 μm thick serial sections. Immunohistochemistry applied to individual sections produces a 2-dimensional survey of the developing hESC colony. Based on serial paraffin sections of the 2D-expression pattern, a new and useful 3D-visualization can be modeled. The actual 3D rendering of an entire colony is accomplished using 3D image processing software such as Mimics(®) or Amira(®). An extended version of this technique even allows for a high-magnification 3D-reconstruction of, e.g., hepatic stem cells in developing liver. These techniques combined allow for both a 2- and a 3-dimensional visualization of hESC colonies and stem cells in organs, which leads to new insights into and information about the interaction of stem cells with their surroundings.

OriginalsprogEngelsk
TitelHuman Embryonic Stem Cell Protocols
Antal sider18
Vol/bind1307
ForlagSpringer Science+Business Media
Publikationsdato2015
Sider245-262
ISBN (Trykt)978-1-4939-2667-1
ISBN (Elektronisk)978-1-4939-2668-8
DOI
StatusUdgivet - 2015
NavnMethods in Molecular Biology
Vol/bind1307
ISSN1064-3745

ID: 162378790